Academic Sciences International Journal of Pharmacy and Pharmaceutical Sciences

ISSN- 0975-1491 Vol 5, Issue 1, 2013

Research Article

IN VITRO ANTIMICROBIAL, ANTIOXIDANT, ANTIARTHRITIC AND PHYTOCHEMICAL

EVALUATION OF PSCYCHOTRIA FLAVIDA TALBOT, AN ENDEMIC PLANT OF WESTERN GHATS

SANA SHEIK, K.R.CHANDRASHEKAR

Department of Applied Botany, Mangalore University, Mangalagangothri- 574199, Konaje, India. Email: konambi @yahoo.com
Received: 19 Oct 2012, Revised and Accepted: 30 Nov 2012

ABSTRACT
The present study deals with the investigation on the in vitro antimicrobial, antioxidant, and antiarthritic activity in the dried extracts of leaf and
stem of Pscychotria flavida Talbot. The study revealed that plant contains several physiologically active phytochemicals such as phenols, saponins,
flavonoids, tannins, steroids and glycosides. Crude methanol, ethyl acetate and aqueous extracts of the leaf and stem were evaluated for
antimicrobial activity by disc diffusion method, antioxidant activity by DPPH and reducing power assay, antiarthritic activity by anti protein
denaturation and proteinase inhibition method. A high linear correlation between phenols and DPPH antioxidant activity/reducing power assay
was observed. Flavonoids correlated only with reducing power. Methanol extract of both leaf and stem exhibited higher antibacterial activity which
is correlated to the presence of higher amount of phenols. Methanol extract of leaf and ethyl acetate of stem showed on par antifungal activity
against Candida albicans which is comparable to that of standard Nystatin. The extract also showed significant inhibition of protein denaturation
and proteinase inhibition.
Keywords: Pscychotria flavida, Phytochemicals, Antimicrobial, Antiarthritic, Antioxidant.

INTRODUCTION Pscychotria flavida Talbot, belonging to the Family Rubiaceae is an

Medicinal plants, which form the backbone of traditional medicine,
have in the last few decades been the subject of very intense
pharmacological studies. Plant derived drugs serve as a prototype to
develop more effective and less toxic medicines [1]. The curative
properties of medicinal plants are perhaps due to the presence of
various secondary metabolites such as alkaloids, flavonoids, phenols,
saponins, sterols etc [2]. There is a growing attention in correlating
the phytochemicals of a medicinal plant with its pharmacological
activity [3].
endemic shrub commonly known as South Indian Wild Coffee
abundantly found in the evergreen forests of Western Ghats and
West Coast. It is used as medicines for wound healing among tribal
people in Southern India. Root is dried, powdered, mixed with
coconut oil and applied topically on affected places to treat wounds
[10]. Traditional use in medicine and phytochemical compounds of
pharmacological interest of Pscychotria flavida Talbot has instigated
the investigations for possible biological activities.
MATERIALS AND METHODS

Pathogens have evolved numerous defence mechanisms against Collection of material

antimicrobial agents, and resistance to old and newly produced
drugs is on the rise. The increasing failure of chemotherapeutic
agents and antibiotic resistance exhibited by pathogenic microbial
infectious agents has led to the screening of several medicinal plants
for their potential antimicrobial activity [4].
Medicinal plants typically contain mixtures of different chemical
compounds that may act individually or in synergy to improve the
health of common man. Antioxidant supplements rich in plants
may be used to help the human body in reducing the oxidative
damage by free radicals. Since the imbalance between antioxidants
and free radicals leads to oxidative stress which may result in
tissue injury and subsequent diseases such as atherosclerosis,
heart failure, neurodegenerative disorders, ageing, cancer,
diabetes mellitus, hypertension etc., the utilization of effective
antioxidants of natural origin is desired [5]. It has been proved
that certain non-nutritive chemicals in plants such as terpenoids,
flavonoids, phenolic compounds which were earlier thought to be
of no importance to human diet, possess antioxidant properties
[6]. In addition to vitamin C, polyphenols (phenolic acids,
catechins, flavonols and anthocyanins), the natural antioxidants
obtained from plants are of greater benefit in comparison to
synthetic ones. Most frequently used synthetic antioxidants in food
industry at high doses, such as BHA, exhibit genotoxic and
carcinogenic effect [7].
Free radicals are important mediators that provoke inflammatory
processes of which consequently, their neutralization by
antioxidants and radical scavengers can attenuate inflammation [8].
Rheumatoid arthritis is a chronic, systemic inflammatory disease
predominantly affecting the joints and peri-articular tissues. The
screening and development of drugs for their anti-inflammatory
activity is still in progress and there is hope for finding anti-
inflammatory drugs from indigenous medicinal plants [9].
Healthy leaf and stem samples were collected from the plants located
at Charmadi forests of Western Ghats during the month of April 2012.
The collected samples were shade dried, ground into fine powder
using domestic grinder and stored in sterile polythene bags until use.
Preparation of extract
Fifty grams of the powdered leaf and stem samples were soxhleted
separately using methanol, and ethyl acetate as solvents for 48
hours. The extracts obtained were evaporated to dryness. Aqueous
extracts were prepared by soaking the samples in water for 72 hours
in a water bath and the obtained extracts were filtered through 6
layers of muslin cloth, centrifuged at 5000g for 15 minutes and the
collected supernatant was evaporated to dryness. All extracts were
stored at 4°c for further use.
Phytochemical analysis
Preliminary screening was performed in all the extracts for
phytochemicals such as alkaloids (Hagers, Wagners, Mayers,
Dragendorff’s tests), flavonoids (Shinodas test), steroids
(Libermann- Burchard and Salkowski tests), phenols (FeCl3 test),
tannins (lead acetate test), saponins (foam test), glycosides
(Molisch’s, sodium hydroxide test) resins (turbidity test) [11].
Determination of Total Phenolic content
The total phenolic content was measured using the Folin-Ciocalteau
method12. Hundred μl aliquot of stock sample (10 mg/ml) was mixed
with 2.0 ml of 2% Na2CO3 and allowed to stand for 2 min at room
temperature. Then 100 μl of 50% Folin Ciocalteau’s phenol reagent
was added. After incubation for 30 min at room temperature in
darkness, the absorbance was read at 720nm using
spectrophotometer. The total phenolic contents of the samples were
expressed as mg gallic acid equivalent per gram of extract (mg GAE/g).
 Chandrashekar et al.
Determination of Flavonoid content
Total flavonoid content was determined following the Aluminium
chloride method [13]. A known aliquot of each extract was made up
to 4 ml using distilled water and 0.3 ml of NaNO2 ( 1:20) was added.
After 5 min, 0.3 ml of 10% AlCl3.H2O solution was added. After 6 min,
2ml of 1 M NaOH solution was added and then the total volume was
made up with 2.4 ml distilled water. The absorbance against blank
was determined at 510 nm. Results were expressed as mg Quercetin
equivalents (QE)/g of extract.
Antimicrobial activity by disc diffusion method
Six bacterial cultures viz. two Gram-positive (Staphylococcus aureus
NCIM 2079, Bacillus subtilis ATCC 6633), four Gram-negative
(Escherichia coli NCIM 2931, Kliebsiella pneumoniae NCIM 2957,
Proteus vulgaris NCIM 2813 and Pseudomonas aeruginosa NCIM
2200) were obtained from National chemical laboratory, Pune, India
and were maintained on nutrient agar slants. Two hundred μl of
overnight grown culture of each organism was dispensed into 20 ml
of sterile nutrient broth and incubated for 4-5 hrs at 37oC to
standardize the culture to 10-5CFU/ml. The fungal strains viz.
Aspergillus niger MTCC No. 1344, Candida albicans MTCC No. 227
were obtained from IMTECH, Chandigarh, India and Trichoderma
viridae was obtained from Plant pathology laboratory CPCRI,
Kasaragod, India.
Antibacterial and antifungal assay were carried out by disc diffusion
method. For this, 0.1ml (10-5CFU /ml) of 24 hrs old bacterial culture
was placed on Muller Hinton agar medium and spread throughout
the plate by spread plate technique. Sterile paper discs (6mm in
diameter) impregnated with 25 μl of the extract (35mg/ml) was
placed on the surface of the medium and incubated at 37oC for
24hrs. Antibacterial activity was recorded by measuring the
diameter of zone of inhibition. Streptomycin was used as positive
reference standard. The antifungal activity was assayed by
inoculating the fungal spores on the potato dextrose agar (PDA)
medium discs pre-impregnated with plant extracts. For Candida
albicans, inoculum was prepared by taking 5-8 colonies of fungal
strain from the fresh cultures and suspended in 5 ml of sterile
distilled water. Nystatin was used as positive standard against
fungal strains.
Percentage inhibition = 100- {(Optical density of test solution – Optical density of product control) ×100}
(Optical density of test control)
The control represents 100% protein denaturation. The results were
compared with standard.
Proteinase inhibitory action [17]
A mixture of 1.0 ml of 25 mM tris-HCl buffer (pH 7.4),100 μg of
extract in 1ml of water and 0.06mg of trypsin was incubated at 37°C
for 5 minutes. Then, 1.0 ml of 0.8% (w/v) casein was added. The
mixtures were incubated for an additional 20 minutes. 2.0 ml of 70%
(v/v) perchloric acid was added to terminate the reaction. The
cloudy suspension was centrifuged and the absorbance of the
supernatant was read at 280 nm (UV- 1800 Shimadzu
spectrophotometer) against buffer as blank. The percentage of
proteinase inhibitory action was calculated as follows:
Percentage inhibition =100 – {(optical density of sample) ÷ (optical
density of control) × 100}.
The results were compared with Diclofenac sodium (100μg/ml).
Statistical Analysis
All the experiments were performed in triplicates (n=3). Statistical
analysis was carried out using Graph Pad Prism Software. Statistical
differences between extract activities were determined using one
way ANOVA with Bonferroni test. Differences were considered
statistically significant when p < 0.05.
RESULTS
The percentage yield of extract obtained for methanol, water and
ethyl acetate in leaf was 37.69, 25.77, and 15.16 and in stem it was
27.64, 13.31, and 11.554 respectively.
Int J Pharm Pharm Sci, Vol 5, Issue 1, 214-218
Evaluation of DPPH scavenging activity [14]
A solution of DPPH (0.135 mM) in methanol was prepared and 1 ml
of this solution was mixed with1ml of varying concentrations of the
extracts. The reaction mixture was vortexed thoroughly and left in
dark at room temperature for 30 minutes. The absorbance of the
mixture was measured at 517 nm using Ascorbic acid as standard.
The ability to scavenge DPPH radical was calculated as:
% DPPH radical scavenging activity= (Absorbance of control- Absorbance of sample) X 100
(Absorbance of control)
The activity was expressed as 50% inhibitory concentration (IC50)
based on the percentage of DPPH radicals scavenged. Lower the IC50
value, higher is the antioxidant activity.
Reducing power assay [15]
Hundred µl of the extracts of varied concentrations were mixed with
phosphate buffer (2.5 ml, 0.2 M, pH 6.6) and 1% potassium
ferricyanide (2.5 ml). The mixture was incubated at 50°C for 20 min.
2.5 ml of 10% trichloroacetic acid was added to the mixture and
centrifuged at 3000 rpm for 10 min. The supernatant (2.5 ml) was
mixed with distilled water (2.5 ml) and freshly prepared FeCl3
solution (0.5 ml, 0.1%). The absorbance was measured at 700 nm.
Reducing power was expressed as ascorbic acid equivalent (AAE) in
milligram per gram of extract.
In vitro anti-arthritic activity by protein denaturation inhibition
method [16]
The solutions used were the test solution (0.5ml) consisting of plant
extract of 0.05ml (250μg/ml) and bovine serum albumin of 0.45ml
(5%w/v aqueous solution); the test control solution (0.5ml)
consisting of 0.45ml bovine serum albumin and 0.05ml of distilled
water; the product control solution (0.5ml) consisting of 0.45ml of
distilled water and 0.05ml of plant extract and the standard solution
(0.5ml) consisting of 0.45ml of Bovine serum albumin and 0.05ml of
Diclofenac sodium (250μg/ml). Then pH was adjusted to 6.3 using
1N HCl and incubated at 37ºC for 20 min and later at 57ºC for 3min.
After cooling the solutions, 2.5ml of phosphate buffer was added,
and then the absorbance was measured at 416nm. The percentage
inhibition of protein denaturation was calculated as
Phytochemical screening
The phytochemical screening showed the presence of phenols,
flavonoids, glycosides, and tannins in all the extracts assessed
(Table 1). Saponin was present in methanol extract of leaf and
water extracts of both leaf and stem. Alkaloids and resins were not
detected in any of the extracts. Steroids were present in methanol
and ethyl acetate extracts of both leaf and stem.
Total phenolics and flavonoid content
In screening of phenolics there was a wide variation in the amount
which ranged from 36.9 to 365.58 mg GAE/g. The highest phenolic
content (365.88 mg GAE/g) was exhibited in ethyl acetate extract of
stem followed by the methanol extract of leaf sample (250.62 mg
GAE/g) while water extract of both stem and leaf showed lower
phenolic content. The flavonoid content in terms of quercetin
equivalent was between 9.0 to 86.4mg QE/g of the material, the
highest being 86.4mg QE/g in ethyl acetate extract of stem followed
by 42.41 mg QE/g in methanol extract of stem.
Antioxidant activity
Significantly on par IC50values of 7.6, 7.33, 7.13 and 10.6 µg/ml for
DPPH activity were observed in methanol leaf, ethyl acetate leaf,
ethyl acetate stem and methanol stem respectively. The standard
ascorbic acid IC50value was 6.53 µg/ml. The significantly highest
reducing power of 515.35 mg AAE/g for ethyl acetate stem extract
followed by 420.55 mg AAE/g for methanol leaf extract were
observed in the present study.
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Int J Pharm Pharm Sci, Vol 5, Issue 1, 214-218

Table 1: Qualitative Phytochemical Screening

Methanol Water Ethyl acetate
Phytochemicals Leaf Stem Leaf Stem Leaf Stem
Phenols + + + + + +
Flavonoids + + + + + +
Tannins + + + + + +
Saponins + _ + + _ _
Alkaloids _ _ _ _ _ _
Glycosides + + + + + +
Steroids + + _ _ + +
Resins _ _ _ _ _ _
+ Detected, - Not detected.
Table 2: Anti arthritic activity in Pscychotria flavida
Extracts Leaf Stem
Methanol Ethyl acetate Water Methanol Ethyl acetate Water DS
Inhibition of protein 73±5.19 62.66± 5.13 66.66±1.52 67.33±1.52 63.33±5.1 3 62.33± 2.08 85±3.60
Denaturation 250µg/ml
Proteinase inhibition 100µg/ml 69.03±5.71 65.28±4.21 64.18±1.16 66.02±3.92 66.74±3.68 58.64± 0.70 66.82±4.26
Results with the same alphabets are not significant. DS- Diclofenac sodium
[

Antiarthritic activity significantly lower than the standard Streptomycin. However, on

Significantly on par anti arthritic activity in different extracts of P.
flavida and standard Diclofenac sodium was observed in terms of
inhibition of protein denaturation (250µg/ml) and proteinase
inhibition (100µg/ml).
Antimicrobial activity
All the extracts exhibited diverse degrees of antibacterial activity
against the tested bacteria with broad spectrum activity. The
antibacterial activity of all the extracts of P. flavida was
par higher antibacterial activity of methanol extracts of leaf and
stem was observed against S. aureus. The methanol extract of leaf
showed next highest activity against K. pneumoniae. The moderate
antibacterial activity of leaf and stem extracts were against B.
subtilis and E.coli. Lowest activity was observed against P.vulgaris.
Among the extracts tested for antifungal activity, leaf methanol
extract and stem ethyl acetate extract exhibited significantly on
par activity with that of standard Nystatin against C. albicans
(Table3). There was no antifungal activity observed against
A.niger, T.harzianum.

Table 3: Antimicrobial activity

Extract Zone of Inhibition (diameter in mm)
TV AN CA BS SA KP EC PA PV
ML ─ ─ 24.26±0.30a 14.86±0.11a 17.86±0.23a 16.7±0.3 15.33±0.57 12.1±0.17a 9.6±0.15a
WL ─ ─ 10.1±0.1b 12.56±0.15b 14.4±0.36b 11.3±0.36a 13.26±0.46ab 9.7±0.1b 9±0.1b
EL ─ ─ 12.53±0.25b 14.43±0.30a 16.23±0.25 12.3±0.60a 13.93±0.11a 11.1±0.17ab 9.56±0.32ab
MS ─ ─ 20 14.86±0.15a 17.66±0.30a 11.83±0.15a 10.26±0.37c 10.66±0.1b 9.13±0.15b
WS ─ ─ 10.43±0.40b 14.46±0.37a 14.46±0.37b 7.53±0.50 10.1±0.1c 9.63±0.20b 9.03±0.05b
ES ─ ─ 25.06±0.11a 12.1±0.17b 14.33±0.57b 11.06±0.20a 12.23±0.2b 14.5±0.3 10.43±0.05
Streptomycin 22.16±0.37 25.53±0.32 24.1±0.1 19.13±0.20 20.63±0.15 30.1±0.1
Nystatin 16.23±0.25 18.9±0.32 25.23±0.20a
Results with the same alphabets in each column are not significant. TV-Trichoderma viridae, AN- Aspergillus niger, CA- Candida albicans, BS-Bacillus
subtilis, SA-Staphylococcus aureus, KP-Kliebsiella pneumoniae, EC- Escherichia coli, PA-Pseudomonas aeruginosa, PV- Proteus vulgaris. L-Leaf, S-Stem,
M-Methanol, E-Ethyl acetate, W-Water, ─ no activity.

DISCUSSION DPPH assay reaction depends on the ability of the samples to

The presence of steroids, triterpenoids and flavonoids in qualitative
phytochemical screening of ethyl acetate extract of Mussaenda
erythrophylla stem has been reported [18]. Phytochemicals were
strongly present in the ethanol and methanol extracts than the water
extract in heartwood of Tecoma stans [19]. Similar results were
observed in the present study also, where methanol and ethyl acetate
extracted more components than water.Total phenolic content of
154.81mgGAE/g in the water extracts and 169.06 mg GAE /g in
methanol extract leaves of Teucrium montanum L. var. montanum,
f.supinum (L.) Reichenb of leaves was reported 7. The phenol content of
P.flavida was comparatively higher than this report. However, authors
have reported a higher phenolic content of 412.8 mg GAE/g extract in
methanolic extract of stem bark of Erythrina indica [20]. Phenols are
very important plant constituents because of their scavenging ability
on free radicals due to their hydroxyl groups.Therefore, the phenolic
content of plants may contribute directly to their antioxidant action
[21].The leaf ethyl acetate extract of Teucrium montanum had
flavonoid content of 58.48 mg RU/g7. Flavonoids are potent
antioxidants and free radical scavengers, which prevent oxidative cell
damage and have strong anti-cancer activity [22].
scavenge free radicals which is visually noticeable as the colour
change from purple to yellow due to hydrogen donating ability [23].
The more rapid the absorbance decrease, the more potent the
primary antioxidant activity [24]. In reducing power assay the
antioxidant in the extract reduces ferricyanide complex to the
ferrous form. The ethyl acetate extract of Hydnophytum formicarum
Jack had shown high radical scavenging activity with IC50 8.40 µg/ml
in DPPH assay3. A reducing power of 531.75 mg AAE/g in Ocimum
basilicum is reported [25].
There was a positive linear correlation between phenolic content
and DPPH (R2=0.7979) as well as reducing power (R2= 0.9589) (Fig
1), which indicates the significant contributions of phenols to
antioxidant activity. Flavonoids were not significantly correlated
with DPPH (R2 = 0.5012) but reasonably correlated with reducing
power (R2= 0.7285) (Fig 2). Negative correlation between DPPH
activity and flavonoids was reported earlier [26]. It is known that
only flavonoids with a certain structure and particularly hydroxyl
position in the molecule can act as proton donating and show radical
scavenging activity.

216
 Chandrashekar et al.
Fig. 1: Linear correlation between (a) phenols and DPPH (b) phenols and reducing power
Fig. 2: Linear correlation between (a) flavonoids and DPPH (b) flavonoids and reducing power
Phenols are known to be toxic for micro organisms. Flavonoids are
found to be effective due to their ability to complex with
extracellular and soluble proteins and also disruption of microbial
membrane [27].While studying the antimicrobial activities of
methanol extract of Argyreia argentea stem, the authors observed 14
mm zone of inhibition against both gram positive bacteria B. subtilis
and gram negative bacteria E. coli whereas no inhibition was
observed against Aspergillus niger [28]. More or less similar results
were observed in the present study.
Abutilon indicum (Linn.) Sweet. at a concentration of 250µg/ml
provided 62.72% protection against denaturation of proteins and at
a concentration of 100µg/ml exhibited 60.21 % antiproteinase
activity [29]. A slightly higher inhibition of protein denaturation and
proteinase inhibition was observed in the present study. The
production of auto antigen in certain arthritic diseases may be due
to the denaturation of protein 9. This effect may be due to the
presence of steroids, alkaloids and flavonoids. The mechanism of
denaturation involves alteration of electrostatic hydrogen,
hydrophobic and disulphide bonding [30]. Proteinases has also been
implicated in arthritic reactions. It was previously reported that
leucocytes proteinases play an important role in the development of
tissue damage during inflammatory reactions and significant level of
protection was provided by proteinase inhibitors [29].
Comparatively good proteinase inhibition activity exhibited by
different extracts of Pscychotria flavida is expected to protect protein
denaturation possibly by controlling the production of autoantigens.
Further research on these lines may throw light on the possible
mechanism of action.
Phytochemical analysis revealed the presence of phenols, flavonoids,
tannins and saponins in Pscychotria laved Talbot. Phytochemical
compounds of pharmacological activity may be responsible for the
antimicrobial, antioxidant and anti denaturation properties.
Methanolic leaf and ethyl acetate stem extract showed high phenolic
compounds, reasonable antimicrobial and antioxidant activity.
Therefore, Pscychotria flavida can be used as a source of potent drug
by mankind for its therapeutic value.
Int J Pharm Pharm Sci, Vol 5, Issue 1, 214-218
ACKNOWLEDGEMENT
Senior author (Sana) is grateful to DST for awarding the INSPIRE
fellowship. Facilities provided by the Department of Applied Botany,
Mangalore University are gratefully acknowledged. The authors are
grateful to Dr Soorya Prakash Shenoy for taxonomical identification.
REFERENCES
1. Murugan P, Rajesha A, Athiperumalsami T, Mohan VR.
Screening of Certain Ethnomedicinal Plants for Antibacterial
Activity. Ethnobotanical Leaflets 2008; 12: 433-438.
2. Mallikharjuna PB, Rajanna LN, Seetharam YN, Sharanabasappa
K. Phytochemical studies of Strychnos potatorum L.f.-A
Medicinal Plant. E-J.Chem 2007; 4: 510-518.
3. Prachayasittikul S, Buraparuangsang P, Worachartcheewan A,
Isarankura-Na-Ayudhya C, Ruchirawat S, Prachayasittikul V.
Antimicrobial and antioxidant activity of bioreactive
constituents from Hydnophytum formicarum Jack. Molecules
2008; 13: 904–921.
4. Sharma A, Patel VK, Rawat S, Ramteke P, Verma R.
Identification of the antibacterial component of some Indian
medicinal plants against Klebsiella pneumoniae. Int J Pharm
Pharm Sci 2010; 2(3):123-127.
5. Arunachalam K. Antioxidant and antimicrobial potential of
methanolic extract of Indian sacred grove Gymnostachyum
febrifugum Benth. root. Int J Pharm Biomed Res 2011; 2(3): 67-
71.
6. Tapan Seal. Antioxidant Activity of Some Wild Edible Fruits of
Meghalaya State in India. Advan. Biol. Res 2011; 5 (3): 155-160.
7. Milan S, Stankovic, Neda N, Topuzovic M, Solujic S. Total Phenolic
content, flavonoid concentrations and antioxidant activity, of the
whole plant and plant parts extracts from Teucrium Montanum L.
Var. Montanum, F. Supinum (L.)Reichenb. Biotechnol. &
Biotechnol. Eq 2011. 25(1): 2222-2227.
8. Chatterjee A. The Treatise of Indian Medicinal Plants, National
Institute of Science and Communication CSIR, New Delhi 1997;
IV: 212-217.
217
 Chandrashekar et al.
9. Lavanya R, Maheshwari S, Harish G, Bharath J, Kamali S,
Hemamalani D, Bharath Varma J, Umamaheswara C R.
Investigation of In-vitro anti-Inflammatory, anti-platelet and
anti-arthritic activities in the leaves of Anisomeles malabarica
Linn. Res. J. Pharm., Biol. Chem. Sci 2010; 1(4)
10. Ayyanar M, Ignacimuthu S. Herbal medicines for wound healing
among tribal people in Southern India: Ethnobotanical and
Scientific evidences. Intl J Appl Res Nat Prod 2009; 2(3): 29-42.
11. Dey PM, Harborne JB. Methods in Plant Biochemistry. London:
Academic Press; 1987.
12. Taga MS, Miller EE, Pratt DE. Chia seeds as a source of natural
lipid antioxidants. J. Am. Oil Chem. Soc. 1984; 61:928-931.
13. Zishen J, Mengcheng T. and Jianming, W. The determination of
flavonoid contents in mulberry and their scavenging affects on
superoxide radicals. Food Chem 1999; 64: 555-559.
14. Liyana- Pathiranan CM, Shahidi F. Antioxidant activity of
commercial soft and hard wheat (Triticum aestivum L) as
affected by gastric pH conditions. J. Agri. Food Chem 2005; 53:
2433-2440.
15. Oyaizu M. Studies on product on browning reaction prepared
from glucose amine. Jpn. J. Nutr 1986; 44: 307-315.
16. Mizushima Y, Kobayashi M. Interaction of anti-inflammatory
drugs with serum proteins, especially with some biologically
active proteins. J. Pharm. Pharmacol 1968; 20: 169-173.
17. Seema CC, Meena V. Antioxidant, an anti-inflammatory and
anti-arthritic activity of Centella asiatica extracts J. Chem. Bio.
Phy. Sci 2011; 1 (2): 260– 269.
18. Eswaraiah MC, Elumalai A. Isolation of phytoconstituents from
the stems of Mussaenda erythrophylla. Der Pharmacia Sinica
2011; 2 (6):132-142.
19. Muthu AK, Borse L, Thangatripathi A, Borse SL. Antimicrobial
activity of Heartwood of Tecoma stans. Int J Pharm Pharm Sci
2012; 4 (3).
20. Sowndhararajan K, Joseph M, Rajendrakumaran D. In vitro
xanthine oxidase inhibitory activity of methanol extracts of
Int J Pharm Pharm Sci, Vol 5, Issue 1, 214-218
Erythrina indica Lam. leaves and stem bark. Asian Pac J Trop
Biomed 2012; 10
21. Tosun M, Ercisli S, Sengul M, Ozer H, Polat T. Antioxidant
Properties and Total Phenolic Content of Eight Salvia Species
from Turkey. Biol Res 2009; 42(2): 175-181.
22. Okwu DE, Josaiah C. Evaluation of the chemical composition of
two Nigerian medicinal plants. Afr. J. Biotechnol 2006; 5(4):
357-361.
23. Jamuna KS, Ramesh CK, Srinivasa TR and Raghu Kl. In-vitro
antioxidant studies of some fruits.Int J Pharm Pharm Sci 2011;
3(1):60-63.
24. Saumya SM and Mahaboob Basha P. In-vitro evaluation of free
radical scavenging activities of Panax ginseng and
Lagerstroemia speciosa: a comparative analysis. Int J Pharm
Pharm Sci 2011; 3(1):165-69.
25. Kiendrebeogo M, Ahmed YC, Roger CH, Zeba B, Charles E,
Lamien A, Odile GN. Antiacetylcholinesterase and antioxidant
activity of essential oils from six medicinal plants from Burkina
Faso. Rev. bras. farmacogn 2012; 21(1): 63-69.
26. Mohammed FG, Nagendra PK, Kong KW, Amin I. Flavonoid,
hesperidine, total phenolic contents and antioxidant activities
from Citrus species. Afr. J. Biotechnol 2010; 9(3): 326-330.
27. Tsuchiya H, Sato M, Miyazaki T, Fujiwara S, Tanigaki S, Ohyama
M, Tanaka T, Iinuma M. Comparative study on the antibacterial
activity of phytochemical flavanones against methicillin resistant
Staphylococcus aureus. J. Ethnopharmacol. 1996; 50: 27-34.
28. Rahman M A, Ahsan T, Islam S. Antibacterial and antifungal
properties of the methanol extract from the stem of Argyreia
argentea. Bangladesh J Pharmacol 2010; 5: 41-44.
29. Deshpande V, Jadhav VM, Kadam VJ. In vitro anti-arthritic
activity of Abutilon indicum (Linn.) Sweet. J Pharm Res 2009;
2(4): 644-645.
30. Shravan KN, Kishore G, Siva KG, Sindhu P. In vitro Anti
inflammatory and anti arthritic activity of Leaves of Physalis
angulata L. Int. J. Pharm & Ind. Res 2011; 1.
218